Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains
that model human diseases. However, production of numerous cloned disease model pigs
by SCNT for large-scale experiments is impractical due to its complexity and
inefficiency. In the present study, we aimed to establish an efficient procedure for
proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear
factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and
treated with insulin to allow for normal growth to maturity, at which point
epididymal sperm could be collected for cryopreservation. In vitro
fertilization and intrafallopian insemination using the cryopreserved epididymal
sperm resulted in diabetes model transgenic offspring. These results suggest that
artificial reproductive technology using cryopreserved epididymal sperm could be a
practical option for proliferation of genetically modified disease model pigs.
Mesenchymal stem/stromal cells (MSCs) are known to be useful for treating local bone diseases. However, it is not known if MSCs are effective for treating systemic bone diseases, as the risk for mortality following intravenous MSC administration has hindered research progress. In this study, we compared the safety and efficacy of intra-bone marrow and intravenous administration of MSCs for the treatment of ovariectomy- (OVX-) induced osteoporosis. Cells capable of forming bone were isolated from the murine compact bones and expanded in culture. Relatively pure MSCs possessing increased potential for cell proliferation, osteogenic differentiation, and inhibition of osteoclastogenesis were obtained by magnetic-activated cell sorting with the anti-Sca-1 antibody. Sca-1-sorted MSCs were administered to OVX mice, which were sacrificed 1 month later. We observed that 22% of the mice died after intravenous administration, whereas none of the mice died after intra-bone marrow administration. With respect to efficacy, intravenous administration improved bone mineral density (BMD) by increasing bone mineral content without affecting bone thickness, whereas intra-bone marrow administration improved BMD by increasing both bone mineral content and bone thickness. These results indicate that intra-bone marrow administration of pure MSCs is a safer and more effective approach for treating osteoporosis.
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